Cephaibols: novel antiparasitics from Acremonium tubakii process for their production, and use thereof

ABSTRACT

The invention relates to compounds of the formula I 
     
       
         AcPhe-Aib-Aib-Aib-x-w-Leu-y-Aib-Hyp-Gln-z-Hyp-Aib-Pro-R  (I) 
       
     
     in which R is Phe-ol or Phe-al and w, x, y, and z have the following meanings: 
     a) w is Gly or Ala; x is Aib and y and z are Iva; 
     b) w is Gly; x, y and z are va; 
     c) w is Gly; x and z are Aib and y is Iva; 
     d) w is Gly; x, y and z are Aib; or 
     e) w is Gly; x and y are Aib and z is Iva; 
     or of the formula II 
     
       
         AcPhe-Iva-Gln-Aib-Ile-Thr-Aib-Leu-Aib-x-Gln-Aib-Hyp-Aib-Pro-Phe-Ser  (II), 
       
     
     wherein x is Hyp or Pro, which are synthesized by  Acremonium tubakii  FH 1685 DSM 12774 during fermentation and released into the culture medium, a process for the isolation of the cephaibols from the culture medium, and their purification, and the use of the cephaibols as pharmacologically active compounds, in particular for the control of parasites.

FIELD OF THE INVENTION

The invention relates to novel peptaibol antibiotics, called cephaibols,which are synthesized from Acremonium tubakii FH 1685 DSM 12774 duringfermentation and released into the culture medium, a process for theisolation of the cephaibols from the culture medium and theirpurification, and the use of the cephaibols as pharmacologically activecompounds, in particular for the control of parasites.

BACKGROUND OF THE INVENTION

Parasitoses are widespread and cause a wide spectrum of pathologicaleffects in humans and animals, which range from slight physiologicaldisturbances to severe, even fatal disorders. Nowadays, many intensivelyresearched parasite disorders are known which threaten the health andthe life of humans, their pets, and farm animals.

Globally, the weakening of the immune system is to be increasingly foundin millions of humans. This group of people is so massively affected byopportunistic parasites that, annually, millions of deaths are mourned.In the age of long-distance travel, even in non-third world countries,which normally have a high standard of hygiene, exotic parasites have tobe expected. This has its origins in the fact that there is an everincreasing trend to go on trekking trips under local hygiene conditionsand that—because of the supposed safety in one's own country—there is aloss of the feeling/knowledge about hygiene dangers in other countries.In addition to the protection of human health, protection of animalsfrom the suffering and pain caused by parasitoses is required either bya cure or, if possible, by prevention. Economic reasons especially cometo bear in farm animal husbandry, where, owing to unfavorable conditionsfor the keeping and feeding of animals (e.g., in certain forms of massanimal husbandry), parasitic disorders occur which contribute to aquantitative loss (reduction in yield of meat, number of eggs, racingpace) or a qualitative loss (quality of meat and wool). The great damagethat is caused by parasitoses in humans and animals makes their controldesirable, if not indispensable, in the interests of health and economy.

Parasites are unicellular or multicellular organisms which residetemporarily or permanently in (endoparasites) or on (ectoparasites)foreign organisms and live at the expense of the host. In humans andanimals, many parasites lead to subacute and also to dramatic disorders.The unicellular parasites include the protozoa, such as, for example,plasmodia (malaria pathogen), trypanosomes (Chagas disease pathogen),amoebae, trichomonads or toxoplasmae. Important unicellular parasitesare the endoparasitic worms (helminths), of which the threadworms(nematodes), tapeworms (cestodes) and leeches (trematodes) can causeserious damage in humans and animals. The multicellular ectoparasitesinclude ticks, mites, lice, fleas and other organisms. Although thereare a large number of agents available for the control and treatment ofparasitic disorders (antiparasitics), on the one hand because of theside effects of these agents and on the other hand because of theincreasing formation of resistance, there is a great need for novel,highly effective protozoicides, anthelminthics, and other parasiticides.Infestation with parasites in particular affects the population of thetropical regions; the number of those affected here is estimated at manyhundreds of millions of people, in addition to the considerable damagein the agricultural field.

The use of chemical substances, whose activity against individualparasites or relatively large groups of parasites is known and which aretoxicologically acceptable in the host (humans, animals) still haveoverriding importance in parasite control.

According to their spectrum of action, a differentiation is made betweenanthelminthics acting against helminths, antiprotozoals active againstprotozoa, insecticides active against insects, and acaricides activeagainst mites (acaria); the last two groups are also summarized underthe term ectoparasiticides.

There has been an increase in pharmaceutical resistance caused by longterm and intensive use, particularly in modern mass animal husbandry.There has also been an increased occurrence of severe side effects topresent medications, in particular of continuous medication of peoplewho, within the framework of progressive globalization, have to work fora relatively long time in the tropics and subtropics. These factorscombined with the high cost of prophylaxis/therapy with certainchemotherapeutics makes the search for inexpensive classes of substanceshaving a different mechanism of action and better tolerabilityessential. Antiparasitics are also needed which are not only effective,but also economically preparable in large amounts, and moreover,environmentally friendly.

Bacteria and fungi via their secondary metabolism by means ofnonribosomal peptide synthetases produce peptides having up to 20 aminoacids, and in some cases structurally unusual amino acids. Many of thepreviously known secondary metabolites having a peptide structurepossess interesting biological actions as antibiotics, enzymeinhibitors, cardiotonics, immunomodulators, insecticides, nematocidesand many others (see, for example, Gräfe, U. Biochemie der Antibiotika,Spektrum Heidelberg, 1992).

Within the structural class of peptide active compounds, the so-calledpeptaibols are distinguished in that they unusually contain many aminoacids (up to 20, among them a high proportion of α-aminobutyric acid(Brückner, H., König, W. A., Greiner, M., Jung, G. Angew. Chem. Int. Ed.Engl. 18:476-477(1979)). In addition, peptaibols are very oftenacetylated at the N terminus and contain a radical having an alcoholgroup (e.g., phenylalaninol) or an aldehyde group at the C terminus.

Examples of peptaibols, as addressed above, are aibellin [J. Antibiotics47:1136-1144 (1994)]; ampullosporin [J. Antibiotics 50:72-728, (1997)];antiamoebin [Structure 6:783-792 (1998)]; clonostachin [J. Antibiotics50:105-110 (1997)]; emerimicins [J. Antibiotics, 27:274-282 (1974)] orzervamicins [J. Antibiotics, 27:321-328 (1974)]. These peptaibols aresynthesized by very different strains of the genera Emericellopsis,Trichoderma, Apiocrea, and many others. They display their antibioticactivity against gram-positive bacteria, against some types of fungi,and against amoebae. The antipyretic and neuroleptic action ofampullosporin has moreover been described.

The previously known peptide active compounds, however, often havedisadvantages that are manifested in unsatisfactory potency of action,high toxicity, and/or undesired side effects.

SUMMARY OF THE INVENTION

The object of the invention is therefore to search for novel microbialpeptide active compounds having improved properties and/or novelmechanisms of action.

This object is achieved, according to the invention, by fermenting thestrain Acremonium tubakii FH 1685 DSM 12774 in a nutrient solutionhaving a carbon and nitrogen source, as well as the customary inorganicsalts, until the novel peptaibols, called cephaibols, accumulate in theculture medium, then isolating the cephaibols from the culture medium,and optionally, separating the cephaibols into the individual activepeptide compounds. The isolated cephaibols are pharmacologically activeand are, thus, suitable for use as pharmaceuticals. Because of theirantiparasitic properties, in particular their strong anthelminticactivity and insecticidal action on ectoparasites, they can be employed,in particular, as antibiotics having action against endo- andectoparasites in animals and in humans.

The invention thus relates to

1. A compound of the formula I (SEQ ID NO:9)

AcPhe-Aib-Aib-Aib-x-w-Leu-y-Aib-Hyp-Gln-z-Hyp-Aib-Pro-R  (I)

wherein R is Phe-ol or Phe-al and w, x, y, and z have the followingmeanings:

a) w is Gly or Ala; x is Aib and y and z are Iva;

b) w is Gly; x, y and z are Iva;

c) w is Gly; x and z are Aib and y is Iva;

d) w is Gly; x, y and z are Aib; or

e) w is Gly; x and y are Aib and z is Iva;

and their physiologically tolerable salts; or

a compound of the formula II (SEQ ID NO:10)

AcPhe-Iva-Gln-Aib-Ile-Thr-Aib-Leu-Aib-x-Gln-Aib-Hyp-Aib-Pro-Phe-Ser  (II)

wherein x is Hyp or Pro; and their physiologically tolerable salts.

2. A process for the preparation of one or more compounds of the formulaI or II, which comprises fermenting Acremonium tubakii, in particularAcremonium tubakii FH 1685 DSM 12774, in a culture medium until one ormore compounds of the formula I or II accumulate in the culture mediumand isolating these from the culture medium.

3. The use of a compound of the formula I or II as a pharmacologicallyactive substance, in particular as an antibiotic against human or animalparasites, preferably against helminths.

4. A pharmaceutical preparation for use in humans and/or in animals,comprising one or more compounds of the formula I or II.

DETAILED DESCRIPTION OF THE INVENTION

In the following, the invention is described in detail, in particular inits preferred embodiments.

Compounds of the formula I or II are also designated as peptide activecompounds or cephaibols.

In the following sequences, AcPhe is N-acetylphenylalanine, Aib isα-aminoisobutyric acid, Ala is alanine, Iva is isovaline, Hyp ishydroxyproline, Phe-ol is phenylalaninol, Phe-al is phenylalaninal, Pheis phenylalanine, Gly is glycine, Leu is leucine, Gln is glutamine, Prois proline, lie is isoleucine, Thr is threonine and Ser is serine.

Cephaibol A designates a compound of the formula I, in which R isPhe-ol, w is Gly, x is Aib and y and z are Iva.

AcPhe-Aib-Aib-Aib-Aib-Gly-Leu-Iva-Aib-Hyp-Gln-Iva-Hyp-Aib-Pro-Phe-ol(SEQ ID No. 1)

Cephaibol A1 designates a compound of the formula I, in which R isPhe-ol, w is Ala, x is Aib and y and z are Iva.

AcPhe-Aib-Aib-Aib-Aib-Ala-Leu-Iva-Aib-Hyp-Gln-Iva-Hyp-Aib-Pro-Phe-ol(SEQ ID No. 8)

Cephaibol B designates a compound of the formula I, in which R isPhe-ol, w is Ala and x, y and z are Iva.

AcPhe-Aib-Aib-Aib-Iva-Gly-Leu-Iva-Aib-Hyp-Gln-Iva-Hyp-Aib-Pro-Phe-ol(SEQ ID No. 2)

Cephaibol C designates a compound of the formula I, in which R isPhe-ol, w is Ala, x and z are Aib and y is Iva.

AcPhe-Aib-Aib-Aib-Aib-Gly-Leu-Iva-Aib-Hyp-Gln-Aib-Hyp-Aib-Pro-Phe-ol(SEQ ID No. 3)

Cephaibol D designates a compound of the formula I, in which R isPhe-ol, w is Ala and x, y and z are Aib.

AcPhe-Aib-Aib-Aib-Aib-Gly-Leu-Aib-Aib-Hyp-Gln-Aib-Hyp-Aib-Pro-Phe-ol(SEQ ID No. 4)

Cephaibol E designates a compound of the formula I, in which R isPhe-ol, w is Ala, x and y are Aib and z is Iva.

AcPhe-Aib-Aib-Aib-Aib-Gly-Leu-Aib-Aib-Hyp-Gln-Iva-Hyp-Aib-Pro-Phe-ol(SEQ ID No. 5)

Cephaibol P designates a compound of the formula II, in which x is Hyp.

AcPhe-Iva-Gln-Aib-Ile-Thr-Aib-Leu-Aib-Hyp-Gln-Aib-Hyp-Aib-Pro-Phe-Ser(SEQ ID No. 6)

Cephaibol Q designates a compound of the formula II, in which x is Pro.

AcPhe-Iva-Gln-Aib-Ile-Thr-Aib-Leu-Aib-Pro-Gln-Aib-Hyp-Aib-Pro-Phe-Ser(SEQ ID No. 7)

The cephaibols according to the invention are produced by Acremoniumtubakii, preferably by Acremonium tubakii FH 1685 DSM 12774. Acremoniumtubakii FH 1685 DSM 12774 possesses a beige-red mycelium and ischaracterized by the conidiophores characteristic of Acremonium species.

An isolate was deposited at the Deutsche Sammlung von Mikroorganismenund Zellkulturen GmbH, Mascheroder Weg 1B, D 38124 Brunswick, Germany,according to the rules of the Budapest Convention, on Mar. 31, 1999,under the following number: Acremonium tubakii FH 1685 DSM 12774.

In a nutrient solution (also called a culture medium) which contains acarbon source and a nitrogen source and the customary inorganic salts,Acremonium tubakii FH 1685 DSM 12774 produces one or more of thecompounds of the formula I or II according to the invention.

Instead of the strain Acremonium tubakii FH 1685 DSM 12774, its mutantsand variants can also be employed which synthesize one or more compoundsof the cephaibols according to the invention. Such mutants can beproduced in a known manner by physical means, for example, irradiation,such as with ultraviolet or X-rays, or chemical mutagens, such as, forexample, ethyl methanesulfonate (EMS), 2-hydroxy-4-methoxybenzophenone(MOB), or N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) or using geneticengineering methods.

Screening for mutants and variants that synthesize one or more of thecephaibols according to the invention is carried out according to thefollowing scheme:

separation of the mycelium after fermentation;

extraction of the mycelium with an organic solvent;

extraction of the cephaibols from the culture filtrate using solidphases or water-immiscible organic solvents;

analysis by means of HPLC, TLC or by testing the biological activity.

The fermentation conditions described below apply to Acremonium tubakii,the deposited isolate Acremonium tubakii FH 1685 DSM 12774, and mutantsand variants of these.

The process according to the invention can be employed for fermentationon a laboratory scale (milliliter to liter range) and on an industrialscale (cubic meter scale). If not stated otherwise, all percentagesrelate to weight. Mixing ratios, in the case of liquids, relate tovolume, if no other details are given.

In a culture medium which contains a carbon source and a nitrogen sourceand the customary inorganic salts, Acremonium tubakii FH 1685 DSM 12774produces compounds of formula I or II according to the invention.

Preferred carbon sources suitable for aerobic fermentation areassimilable carbohydrates and sugar alcohols, such as glucose, lactose,sucrose or D-mannitol and carbohydrate-containing natural products, suchas, for example, malt extract. Possible nitrogen-containing nutrientsare: amino acids, peptides and proteins and their degradation products,such as peptones or tryptones, in addition meat extracts, yeastextracts, ground seeds, for example, of corn, wheat, beans, soybeans orthe cotton plant, distillation residues from alcohol production, meatmeals or yeast extracts, but also ammonium salts and nitrates. Inorganicsalts which the nutrient solution can contain are, for example,chlorides, carbonates, sulfates or phosphates of the alkali or alkalineearth metals, iron, zinc, cobalt and manganese.

The formation of the cephaibols according to the invention proceedsparticularly well in a culture medium that contains approximately 0.1 to5%, preferably 0.3 to 2%, of yeast extract and 0.2 to 5%, preferably 0.5to 3%, of sucrose and 0.1 to 10 g/l, preferably 0.2 to 1.0 g/l, ofmagnesium sulfate and 0.05 to 1.0 g/l, preferably 0.1 to 0.5 g/l, ofpotassium or sodium dihydrogenphosphate and 0.05 to 1.0 g/l, preferably0.1 to 1.0 g/l, of sodium nitrate and 0.01 to 0.1 g/l, preferably 0.02to 0.1 g/l, of potassium chloride and 0,01 to 100 μm, preferably 5 to 20μm, of iron sulfate and traces of zinc sulfate and copper sulfate. Thedetails in % are in each case based on the weight of the entire culturemedium.

In the culture medium, Acremonium tubakii FH 1685 DSM 12774 forms amixture of cephaibols. Depending on the composition of the culturemedium, it is possible for the quantitative proportion of one or more ofthe cephaibols according to the invention to vary. Moreover, thesynthesis of individual cephaibols can be controlled by the compositionof the media, such that one or a number of the cephaibols are notproduced at all or are produced in an amount below the detection limitof the microorganism.

The mixture preferably consists of 8 different, detectable cephaibols(cephaibols A, A1, B, C, D, E and also P and Q). The cephaibols A to Eare preferably formed.

In addition to the cephaibols A1 and A to E (compounds of the formula Iwhich carry phenylalaninol (Phe-ol16) as a characteristic element at theC terminus), compounds of the formula I that carry the aldehydephenylalaninal (Phe-al16) instead of Phe-ol16 as the C terminal groupare also formed in the culture medium of the Acremonium tubakii FH 1685DSM 12774. As a rule, these are minority products to the cephaibols A-Eand are obtained from the culture medium together with these.

The microorganism is cultured aerobically, i.e., for example, withshaking or stirring in shaker flasks or fermenters, and if appropriate,with introduction of air or oxygen. It can be carried out in atemperature range from approximately 18 to 35° C., preferably atapproximately 20 to 30° C., in particular at 22 to 28° C. The pH rangeshould be from about 6 to about 8, preferably from about 6.5 to about7.5. In general, the microorganism is cultured under these conditionsover a period of about 24 to 300 hours, preferably about 36 to 140hours.

Culturing is advantageously carried out in several stages, i.e., one ormore precultures are first prepared in a liquid culture medium, whichare then inoculated into the actual production medium, the main culture,for example, in the volume ratio 1:10. The preculture is obtained, forexample, by inoculating a mycelium into a nutrient solution and allowingit to grow for approximately 36 to 120 hours, preferably 48 to 72 hours.The mycelium can be obtained, for example, by allowing the strain togrow for approximately 3 to 40 days, preferably 4 to 10 days, on a solidor liquid nutrient medium, for example, malt-yeast-agar orpotato-dextrose-agar (standard medium for mold fungi, e.g., from Difco).

The course of the fermentation can be monitored by means of the pH ofthe cultures or of the mycelium volume and by chromatographic methods,such as, for example, thin-layer chromatography or high-pressure liquidchromatography, or testing of the biological activity. The cephaibolsaccording to the invention are obtained both in the mycelium and in theculture filtrate, but the largest part is found in the mycelium.

The isolation procedure described below is used for the purification ofthe cephaibols according to the invention, preferably for thepurification of the cephaibols A, A1, B, C and D.

The isolation or purification of the cephaibols according to theinvention from the culture medium is carried out according to knownmethods taking into account the chemical, physical and biologicalproperties of the natural substances. For testing the peptide activecompound concentration in the culture medium or in the individualisolation steps, it is possible to use thin-layer chromatography, forexample, on silica gel using isopropanol/25% strength NH₃ as an eluantor HPLC. Detection in the case of the thin-layer chromatographicseparation can be carried out, for example, by means of stainingreagents such as anisaldehyde/sulfuric acid, the amount of the substanceformed expediently being compared with a calibration solution. Theby-products of the formula I, in which R is a Phe-al radical, canoptionally be separated from the respective cephaibol A-E by customarypurification methods known to the person skilled in the art, forexample, by chromatography or by recrystallization.

For the isolation of the cephaibols according to the invention, themycelium is first separated from the culture medium by the customaryprocesses and the cephaibols are then extracted from the cell mass usingan optionally water-miscible organic solvent. The organic solvent phasecontains the cephaibols according to the invention; they are optionallyconcentrated in vacuo and further purified as described below.

The culture filtrate is optionally combined with the concentrate of themycelium extract and extracted with a suitable, water-immiscible organicsolvent, for example, with n-butanol. The subsequently separated organicphase is optionally concentrated in vacuo. For the defatting of theproduct of value, it is possible to dilute the concentrate with anonpolar solvent in which the cephaibols according to the invention aresoluble to a very small extent, such as, for example, with hexane,petroleum ether, or diethyl ether. The cephaibols precipitate in thisprocess and the lipophilic impurities remain dissolved and are removedby customary solid/liquid phase separations.

The precipitate, which contains all the cephaibols, is dissolved in 1/30of the original volume of water/methanol. The precipitate dissolvescompletely into solution during the course of this and the solution islyophilized. The lyophilizate, subsequently called the crude product,contains 5 to 50% of cephaibols and is employed for further isolation.

The further purification of one or more of the cephaibols according tothe invention is carried out by chromatography on suitable materials,preferably, for example, on molecular sieves, on silica gel or alumina,on ion exchangers or on adsorber resins or on reversed phases (RP).Using this chromatography, the cephaibols are separated.

The chromatography of the cephaibols is carried out with bufferedaqueous solutions or mixtures of aqueous and organic solutions.

Mixtures of aqueous and organic solutions are understood as meaning allwater-miscible organic solvents, preferably methanol or acetonitrile, ina concentration of 10 to 80% of solvent, preferably 40 to 60% ofsolvent, or otherwise all buffered aqueous solutions which are misciblewith organic solvents.

The separation of the cephaibols on the basis of their differingpolarity is carried out with the aid of reversed phase chromatography,for example, on MCI® (adsorber resin from Mitsubishi, Japan) orAmberlite XAD® (TOSOHMS), on further hydrophobic materials, such as, forexample, on RP-8 or RP-18 phases. Moreover, the separation can becarried out with the aid of normal-phase chromatography, for example,on'silica gel, alumina and the like.

The chromatography of the cephaibols is carried out with buffered oracidified aqueous solutions or mixtures of aqueous solutions withalcohols or other, water-miscible organic solvents. The organic solventsused are preferably propanol and acetonitrile.

Buffered or acidified aqueous solutions are understood as meaning, forexample, water, phosphate buffer, ammonium acetate, citrate buffer in aconcentration of 0.1 mM to 0.5 M, and formic acid, acetic acid,trifluoroacetic acid or all customary acids known to the person skilledin the art, preferably in a concentration of 0.01 to 1%. 0.1% isparticularly preferred.

Chromatography is carried out using a gradient which begins with 100%water and ends with 100% solvent, preferably a linear gradient from 30to 60% propanol or acetonitrile is employed.

Alternatively, it is also possible to carry out gel chromatography orchromatography on hydrophobic phases.

Gel chromatography is carried out on polyacrylamide or mixed polymergels, such as, for example, Biogel-P 2® (Biorad) or Fractogel TSK HW 40®(Merck, Germany, Toso Haas, USA) or Sephadex® (Pharmacia, Sweden).

The sequence of the abovementioned chromatographies is reversible.

A further, very effective purification step for cephaibols iscrystallization. The cephaibols crystallize readily from solutions inorganic solvents and from mixtures of water with organic solvents. Thecrystallization is carried out in a manner known per se, for example, byconcentrating or cooling saturated peptide active compound solutions.

The cephaibols according to the invention are stable in the solid stateand in solutions in the pH range from about 3 to about 8, in particularfrom about 5 to about 7, and can thus be incorporated into customarypharmaceutical preparations.

Physiologically tolerable salts of compounds of the formula I or II areunderstood as meaning both their organic and inorganic salts, such asare described in Remington's Pharmaceutical Sciences (17^(th) Edition,page 1418 (1985)). Because of their physical and chemical stability andsolubility, sodium, potassium, calcium and ammonium salts, inter atia,are preferred for acidic groups; salts of hydrochloric acid, sulfuricacid, phosphoric acid or of carboxylic acids or sulfonic acids, such,as, for example, acetic acid, citric acid, benzoic acid, maleic acid,fumaric acid, tartaric acid and p-toluenesulfonic acid, inter alia, arepreferred for basic groups.

The present invention comprises all stereoisomeric forms of thecompounds of the formula I or II. Asymmetric centers contained in thecompounds of the formula If can all independently of one another havethe S configuration or the R configuration. The invention includes allpossible enantiomers and diastereomers, as well as mixtures of two ormore stereoisomeric forms, for example, mixtures of enantiomers and/ordiastereomers, in all ratios. Enantiomers are thus a subject of theinvention in enantiomerically pure form, both as levorotatory and asdextrorotatory antipodes, in the form of racemates and in the form ofmixtures of the two enantiomers in all ratios. In the presence ofcis/trans isomerism, both the cis form and the trans form and mixturesof these forms in all ratios are a subject of the invention.

The present invention also comprises chemical equivalents of thecompounds of the formula I or II. Equivalents of this type are, forexample, esters, ethers, addition salts, complexes or otherwise partialhydrolysis products.

Because of their valuable pharmacological properties, the cephaibolsaccording to the invention are suitable for use as pharmaceuticals inhuman and/or veterinary medicine. The substances according to theinvention possess pharmacological activity, in particular as anantibiotic against parasites, particularly preferably against endo-and/or ectoparasites.

The present invention thus further relates to the use of the compoundsof the formula I or II according to the invention as human or veterinarypharmaceuticals, in particular as chemotherapeutics against human-and/or animal-pathogenic endo- and/or ectoparasites. The mechanism ofaction of these peptide active compounds is unknown, but a significant,lethal effect against helminths and ectoparasites was detected.

The cephaibols according to the invention are suitable, for example, forthe control of animal- and human-pathogenic trematodes (Fasciolahepatica, Fasciolopsis buski, Fasciola gigantica, Fascioloides magna,Dicrocoelium dendriticum, Opistorchis felineus, Clonorchis sinensis,Paragonimus westermanni, Paragonimus kellikotti, Schistosomahaematobium, Schistosoma japonicum, Schistosoma mansoni) and animal- andhuman-pathogenic nematodes which belong to the family of theTrichuridae, Trichinellidae, Strongyloididae, Ancylostomatidae,Strongylidae, Trichostrongylidae, Metastrongylidae, Oesophagostomatidae,Dictyocaulidae, Protostrongylidae, Angiostrongylidae, Oxyuridae,Ascaridae, Toxocaridae, Dracunculidae, Habronematidae and Filariidae;moreover, the cephaibols according to the invention are suitable for thecontrol of animal- and human-pathogenic ectoparasites which belong tothe arachnids class (family: Argasidae, Ixodidae, Dermanyssidae;Demodicidae, Sarcoptidae, Psoroptidae, Varroidae) and to the insectsclass, which comprise the order of the Phthiraptera (Anoplura,Mallophaga), Diptera and Siphonaptera.

In addition to the antibacterial action, the compounds according to theinvention possess antimycotic, i.e., antifungal properties, includingthe phytopathogenic fungi.

In the case of parasites that have formed resistance to conventionalagents, only novel agents possess a therapeutically adequate action. Thecephaibols of the formula I or II according to the invention thuspotentially have an excellent action even against these problemorganisms.

Because of the antibacterial, antimycotic and antiprotozoal activity,the cephaibols according to the invention possess growth-promotingactions, which can be used to good effect in animal production. Theimproved feed utilization can be utilized in farm animals such as, forexample, cattle, sheep, goats, pigs, horses or rabbits. For this, dosesof 0.05-50 mg/kg/day are preferably used. The cephaibols in this casealso bring about a decrease in the production of methane gas and animprovement in the feed utilization.

The invention also relates to pharmaceutical preparations that containone or more of the cephaibols according to the invention. Use as amixture with suitable excipients or carrier material is preferred.Carrier material which can be used in veterinary pharmaceuticals are thecustomary feedstuff mixtures or, in the case of humans, allpharmacologically tolerable carrier materials and/or excipients.

The invention also relates to a process for the production of apharmaceutical according to the invention, which comprises bringing atleast one of the compounds according to the invention into a suitableadministration form using a pharmaceutically suitable andphysiologically tolerable carrier and, if appropriate, further suitableactive compounds, additives or excipients.

In general, the pharmaceuticals according to the invention areadministered orally, locally, or parenterally, and, in principle rectaladministration is also possible. Suitable solid or liquid pharmaceuticalpreparation forms are, for example, granules, powders, tablets, coatedtablets, (micro)capsules, suppositories, syrups, emulsions, suspensions,aerosols, drops or injectable solutions in ampoule form and alsopreparations with protracted release of active compound, in whoseproduction vehicles and additives and/or excipients, such asdisintegrants, binders, coating agents, swelling agents, glidants orlubricants, flavorings, sweeteners or solubilizers, are customarilyused. Frequently used vehicles or excipients which may be mentioned are,for example, magnesium carbonate, titanium dioxide, lactose, mannitol,and other sugars, talc, lactoprotein, gelatin, starch, vitamins,cellulose and its derivatives, animal or vegetable oils, polyethyleneglycols and solvents, such as, for example, sterile water, alcohols,glycerol, and polyhydric alcohols.

If appropriate, the dose units can be microencapsulated for oraladministration in order to delay the release or to extend it over alonger period of time, such as, for example, by coating or embedding theactive compound in particle form into suitable polymers, waxes, or thelike.

The pharmaceutical preparations are preferably produced and administeredin dose units, each unit containing as active constituent a specificdose of one or more compounds of the cephaibols according to theinvention. In the case of solid dose units such as tablets, capsules andsuppositories, this dose can be up to approximately 2000 mg, butpreferably approximately 1 to 1000 mg, and in the case of injectionsolutions in ampoule form up to approximately 1000 mg, but preferablyapproximately 10 to 300 mg, per day.

The daily dose to be administered is dependent on the body weight, age,sex and condition of the mammal. Under certain circumstances, however,even higher or lower daily doses may be appropriate. The administrationof the daily dose can be carried out either by single administration inthe form of an individual dose unit or in a number of smaller dose unitsand by multiple administration of subdivided doses at specificintervals.

The pharmaceuticals according to the invention are produced by bringingone or more compounds of the cephaibols according to the invention intoa suitable administration form using customary vehicles and, ifappropriate, additives and/or excipients.

In principle, in the treatment of helminths and ectoparasites, adifferentiation is to be made between therapeutic, meta- andprophylactic measures; these different treatment methods requirespecific, pharmaceutical formulations. Formulations of this typeguarantee, for example, the continuous administration of subtherapeuticto therapeutic doses of anthelmintics or of ectoparasiticides, which aredifferentiated into “sustained-” and “pulse-release boli” according totheir mode of release. The former are further differentiated into“slow-release boli”, which are those with a decreasing release rate, and“continuous-release boli”, which are those with a constant release mode.“Pulse-release boli”, on the other hand, release all of the activecompound within a few hours to days. In addition to the releasetechnology, which also comprises the micro-encapsulation of activecompounds and which enjoys increasing popularity in veterinary medicine,so-called “spot on” or “pour on” formulations are customary for externalapplication to the animal. The administration of tablets, pastes, orinjection solutions and the use of neckbands that contain medicamentsagainst, for example, ectoparasites, are known in the prior art.

The invention is illustrated further in the following examples.Percentages relate to weight. Mixing ratios in liquids relate to volume,if no other details have been given.

EXAMPLES Example 1 Preparation of a Glycerol Culture of Acremoniumtubakii FH 1685 DSM 12774

100 ml of nutrient solution (malt extract 2.0%, yeast extract 0.2%,glucose 1.0%, (NH₄)₂HPO₄ 0.05%, pH 6.0) in a sterile 300 ml Erlenmeyerflask are inoculated with the strain Acremonium tubakii FH 1685 DSM12774 and incubated on a rotating shaker for 7 days at 25° C. and 140rpm. 1.5 ml of this culture are then diluted with 2.5 ml of 80% glyceroland stored at −20° C.

Example 2 Preparation of a Culture or a Preculture in the ErlenmeyerFlask of Acremonium tubakii FH 1685 DSM 12774

A sterile 300 ml Erlenmeyer flask containing 100 ml of the followingnutrient solution: 30 g/l of sucrose, 5 g/l of yeast extract, 1 g/l ofK₂HPO₄, 3 g/l of NaNO₃, 0.5 g/l of MgSO₄×7 H₂O, 0.01 g/l of FeSO₄×7 H₂O,0.5 g/l of KCl and 1.0 ml of trace element solution (0.2 g/l of ZnSO₄×7H₂O and 0.7 g/l of CuSO₄×5 H₂O) is inoculated with a culture grown on aslant tube (same nutrient solution, but with 2% agar) or with 1 ml of aglycerol culture (see Example 1) and incubated on a shaker at 180 rpmand 30° C. The maximum production of one or more compounds of thecephaibols according to the invention is achieved after about 120 hours.For the inoculation of 10 and 200 liter fermenters, the 48 to 96hour-old submersed culture (inoculation quantity about 10%) of the samenutrient solution suffices.

Example 3 Preparation of the Cephaibols

A 30-liter fermenter is operated under the following conditions:

Nutrient medium: 30 g/l of sucrose 5 g/l of yeast extract 3 g/l of NaNO₃0.5 g/l of KCl 0.5 g/l of MgSO₄ 0.1 g/l of K₂HPO₄ 10 μM FeCl₃ × 6 H₂O 1ml of trace element solution pH 6.5 (before sterilization) Trace elementsolution: 2 g/l of ZnSO₄ × 7 H₂O and 0.7 g/l of CuSO₄ × 5 H₂O.Incubation time: 50 hours Incubation temperature: 25° C. Stirrer speed:300 rpm Aeration: 15 liter min⁻¹

By repeated addition of ethanolic polyol solution, it is possible tosuppress foam formation. The production maximum is achieved after about96 to 120 hours.

Example 4 Isolation of the Cephaibol Mixture from the Culture Solutionof Acremonium tubakii FH 1685 DSM 12774

After completion of the fermentation of Acremonium tubakii FH 1685 DSM12774, the culture broth from three fermenters, obtained according toExample 3, (90 liters) is filtered with the addition of approximately 2%filter aid (e.g., Celite®) and the cell mass (6 liters) is extractedwith 20 liters of methanol. The peptide active compound-containing,methanolic solution is freed from the mycelium by filtration andconcentrated in vacuo. The concentrate is applied together with theculture filtrate (83 liters) to a previously prepared, 4 liter MCl gel,CHP20P column. This is eluted with a gradient of 0.1% trifluoroaceticacid in water to 0.1% trifluoroacetic acid in propan-2-ol. The flowthrough the column (12 liters per hour) is collected in fractions (2.5liters each) and the fractions containing the peptide active compounds(21 to 24) are combined. Concentration in vacuo and freeze-drying afford4 g of a pale brown powder.

Example 5 Enrichment of the Cephaibol Components by gel Chromatography

4 g of the product obtained according to Example 4 are applied to a 3.9liter capacity column packed with Fractogel® TSK MW-40 s(width×height=10 cm×50 cm). The eluant methanol is pumped through thecolumn at a flow rate of 50 ml per minute and the column efflux iscollected in fractions (65 ml). The cephaibols are found mainly infractions 28 to 33. They are combined and freed from the methanol invacuo. They afford 1.3 g of peptide active compound mixture.

Example 6 Separation of the Cephaibol Components on Reverse-phase RP-18

A 500 ml capacity preparative HPLC column (5.1 cm (ID)×25 cm H) ispacked with ®Nucleosil 100-7 C18 HD and the 1.3 g of the peptide activecompound mixture obtained according to Example 5 are applied. Elution iscarried out with 40% acetonitrile in 0.1% aqueous trifluoroacetic acidsolution. The flow through the column is 50 ml/minute and fractions of acontent of 125 ml each are collected. Cephaibol D is found in fraction46, cephaibol C in fractions 49 and 50, cephaibol E in fraction 51,cephaibol A in fractions 60 to 64 and cephaibol B and A1 in fractions 66and 67. Fraction 68 comprises a mixture of cephaibols. Afterconcentrating in vacuo and freeze-drying, the following amounts areweighed out:

Cephaibol A: 520 mg, ESI+MS: 1671 Da (M+H)⁺,

Cephaibol Al: 4 mg, ESI+MS: 1685 DA (M+H)⁺

Cephaibol B: 38 mg, ESI+MS: 1685 Da (M+H)⁺,

Cephaibol C: 195 mg, ESI+MS: 1657 Da (M+H)⁺,

Cephaibol D: 16 mg, ESI+MS: 1643 Da (M+H)⁺,

Cephaibol E: 76 mg, ESI+MS: 1657 Da (M+H)⁺.

Example 7 Isolation of the Cephaibols P and Q

Fraction 68, obtained according to Example 6, is dissolved, after freezedrying (4.1 mg), in 20% acetonitrile in water and applied to a 250/10Nucleosil C18 300-7® column. Elution is carried out with 0.005% ammoniumacetate buffer in 40% acetonitrile. In addition to a little cephaibol B,1 mg of cephaibol P and 1 mg of cephaibol Q are obtained after drying.

Cephaibol P: ESI+MS a molecular weight of 1873 (M+H )⁺ is measured,

Cephaibol Q: ESI+MS a molecular weight of 1857 (M+H)⁺ is measured.

Example 8 HPLC System for the Detection of the Cephaibols

The system described below allows the separation and quantification ofthe cephaibols in the crude mixture or in the culture filtrate; theretention times are between 7.0 minutes (cephaibol D) and 18.8 minutes(cephaibol A1).

Eluant: 0.1% trifluoroacetic acid in 40% acetonitrile. Column: Nucleosil100 C₁₈AB 250/4, Macherey-Nagel. Flow: 1.0 ml/min. Detection:Ultraviolet light absorption at 210 nm.

Under the conditions indicated, the cephaibols have the followingretention times:

Cephaibol A: 12.1 minutes,

Cephaibol A1: 18.8 minutes,

Cephaibol B: 16.0 minutes,

Cephaibol C: 9.0 minutes,

Cephaibol D: 7.1 minutes,

Cephaibol E: 7.3 minutes,

Cephaibol P: 17.3 minutes,

Cephaibol Q: 17.9 minutes.

Example 9 Characterization of Cephaibol A

10 μg of cephaibol A are hydrolyzed in constant-boiling hydrochloricacid and investigated in an amino acid analyzer. The following customaryamino acids are found:

Hydroxyproline 11.5 nmol  Glutamic acid 5.7 nmol Proline 5.5 nmolGlycine 5.5 nmol Leucine 5.6 nmol Phenylalanine 5.5 nmol

The contents of the unusual amino acids aminoisobutyric acid andisovaline were not assessable.

Mass-spectrometric investigations using a Finnigan MAT LCQ ion-trap massspectrometer with electron spray ionization (ESI+) afford the followingdata:

The ESI mass spectrum shows an intensive MH⁺ at m/e 1670.7 and an[M+Na]⁺ at m/e 1692.7 corresponding to a monoisotopic molecular weightof 1669.7, in good agreement with the calculated mass (forC₈₂H₁₂₇N₁₇O₂₀, monoisotopic) of 1669.9 Da. The MS/MS spectrum shows afragmentation that is listed in bold in Table 1. The fragments printedin italics are calculated, but not observed fragments.

TABLE 1 MS/MS fragmentation of cephaibol A B₁ 190.1 B₁ 212.1 A₁ 184.1 B₂275.1 B₂ 297.1 A₂ 269.1 B₃ 360.2 B₃ 382.2 A₃ 354.2 B₄ 445.2 B₄ 467.2 A₄439.2 B₅ 530.3 B₅ 552.3 A₅ 524.3 B₆ 587.3 B₆ 609.3 A₆ 581.3 B₇ 700.4 B₇722.4 A₇ 694.4 B₈ 799.5 B₈ 821.5 A₈ 793.5 B₉ 884.5 B₉ 906.5 A₉ 878.5 B₁₀997.6 B₁₀ 1019.6 A₁₀ 991.6 B₁₁ 1125.6 B₁₁ 1147.6 A₁₁ 1119.6 B₁₂ 1224.7B₁₂ 1246.7 A₁₂ 1218.7 B₁₃ 1337.7 B₁₃ 1359.7 A₁₃ 1331.7 B₁₄ 1422.8 B₁₄1444.8 A₁₄ 1416.8 B₁₅ 1519.9 B₁₅ 1541.9 A₁₅ 1513.9

The protonated B fragmentation is listed in the first column—and in thesecond column the (M+sodium)⁺-B fragmentation. The third column showsthe A fragment series in the Na form.

The physicochemical and spectroscopic properties of the cephaibolsaccording to the invention can be summarized as follows:

Cephaibol A (SEQ ID NO:1)

Appearance: soluble in polar organic solvents, but only slightly solublein water; colorless substance. It is stable in neutral, mildly acidicand mildly alkaline medium.

Empirical formula: C₈₂H₁₂₇N₁₇O₂₀,

structural formula:

molecular weight: 1671 Da,

UV absorption (λ_(max)): 258 nm, log ε: 2.62;

NMR data: see Table 2.

Cephaibol A1 (SEQ ID NO:8)

Appearance: soluble in polar organic solvents, but only slightly solublein water; colorless substance. It is stable in neutral, mildly acidicand mildly alkaline medium.

Empirical formula: C₈₃H₁₂₉N₁₇O₂₀,

structural formula:

molecular weight: 1685 Da,

UV data (λ_(max)): 258 nm, log ε: 2.62;

NMR data: see Table 3.

Cephaibol B (SEQ ID NO:2)

Appearance: soluble in polar organic solvents, but only slightly solublein water; colorless substance. It is stable neutral, mildly acidic andmildly alkaline medium.

Empirical formula: C₈₃H₁₂₉N₁₇O₂₀,

structural formula:

molecular weight: 1685 Da,

UV absorption (λ_(max)): 258 nm, log ε: 2.62;

NMR data: see Table 4.

Cephaibol C (SEQ ID NO:3)

Appearance: soluble in polar organic solvents, but only slightly solublein water; colorless substance. It is stable in neutral, mildly acidicand mildly alkaline medium.

Empirical formula: C₈₁H₁₂₅N₁₇O₂₀, structural formula:

Molecular weight: 1657 Da,

UV absorption (λ_(max)): 258 nm, log ε: 2.62;

NMR data: see Table 5.

Cephaibol D (SEQ ID NO:4)

Appearance: soluble in polar organic solvents, but only slightly solublein water; colorless substance. It is stable in neutral, mildly acidicand mildly alkaline medium.

Empirical formula: C₈₀H₁₂₃N₁₇O₂₀,

structural formula:

molecular weight: 1643 Da,

UV absorption (λ_(max)): 258 nm, log ε: 2.62;

NMR data: see Table 6.

Cephaibol E (SEQ ID NO:5)

Appearance: soluble in polar organic solvents, but only slightly solublein water; colorless substance. It is stable in neutral, mildly acidicand mildly alkaline medium.

Empirical formula: C₈₁H₁₂₅N₁₇O₂₀,

structural formula:

molecular weight: 1657 Da,

UV absorption (λ_(max)), 258 nm, log ε: 2.62;

NMR data: see Table 7.

Cephaibol P (SEQ ID NO:6)

Appearance: soluble in polar organic solvents, but only slightly solublein water; colorless substance. It is stable in neutral, mildly acidicand mildly alkaline medium.

Empirical formula: C₈₉H₁₃₇N₁₉O₂₅,

structural formula:

molecular weight: 1873 Da,

UV absorption (λ_(max)): 258 nm, log ε: 2.62;

NMR data: see Table 8.

Cephaibol Q (SEQ ID NO:7)

Appearance: soluble in polar organic solvents, but only slightly solublein water; colorless substance. It is stable in neutral, mildly acidicand mildly alkaline medium.

Empirical formula: C₈₉H₁₃₇N₁₉O₂₄,

structural formula:

molecular weight: 1857.197 Da,

UV absorption (λ_(max)): 258 nm, log ε: 2.62;

NMR data: see Table 9.

TABLE 2 NMR data (chemical shifts) for cephaibol A in DMSO at 300° K.Proton/carbon ¹H ¹³C Ac—Me 1.83 22.29 Ac—C′ — 170.37  Phe1-NH 8.29 —Phe1-α 4.34 55.11 Phe1-β 2.97/2.83 36.37 Phe1-γ — 137.37  Phe1-δ 7.29129.16  Phe1-ε 7.29 128.09  Phe1-ξ 7.22 126.39  Phe1-C′ — 172.41 Aib2-NH 8.59 — Aib2-α — 55.77 Aib2-β 1.28 23.57 Aib2-β′ 1.27 25.37Aib2-C′ — 174.92  Aib3-NH 7.64 — Aib3-α — 55.86 Aib3-β 1.32 24.04Aib3-β′ 1.29 24.62 Aib3-C′ — 175.11  Aib4-NH 7.69 — Aib4-α — 55.86Aib4-β 1.39 24.99 Aib4-β′ 1.38 24.55 Aib4-C′ — 175.62  Aib5-NH 7.57 —Aib5-α — 55.92 Aib5-β 1.41 24.99 Aib5-β′ 1.37 24.49 Aib5-C′ — 175.62 Gly6-NH 7.98 — Gly6-α 3.76/3.64 43.43 Gly6-C′ — 170.69  Leu7-NH 7.74 —Leu7-α 4.02 53.18 Leu7-β 1.70/1.53 39.40 Leu7-γ 1.69 24.13 Leu7-δ 0.9222.62 Leu7-δ′ 0.85 21.57 Leu7-C′ — 171.80  Iva8-NH 7.39 — Iva8-α — 59.34Iva8-β 2.20/1.67 27.60 Iva8-γ 0.73  7.27 Iva8-αMe 1.27 21.93 Iva8-C′ —176.07  Aib9-NH 7.57 — Aib9-α — 56.22 Aib9-β 1.48 23.17 Aib9-β′ 1.3725.72 Aib9-C′ — 173.44  Hyp10-α 4.38 61.01 Hyp10-β 2.16/1.77 36.84Hyp10-γ 4.29 68.95 Hyp10-γOH 5.11 — Hyp10-δ 3.73/3.50 56.13 Hyp10-C′ —171.80  Gln11-NH 7.89 — Gln11-α 4.17 52.42 Gln11-β 2.16/1.86 26.66Gln11-γ 2.09 31.44 Gln11-δ — 173.08  Gln11-ε-NH₂ 7.19/6.70 — Gln11-C′ —172.13  Iva12-NH 7.47 — Iva12-α — 58.48 Iva12-β 2.15/1.78 28.06 Iva12-γ0.74  6.97 Iva12-αMe 1.41 20.36 Iva12-C′ — 172.83  Hyp13-α 4.53 60.55Hyp13-β 2.17/1.67 37.33 Hyp13-γ 4.21 69.04 Hyp13-γOH 5.09 — Hyp13-δ3.67/3.37 56.46 Hyp13-C′ — 172.83  Aib14-NH 7.94 — Aib14-α — 55.65Aib14-β 1.40 23.50 Aib14-β′ 1.33 25.66 Aib14-C′ — 171.73  Pro15-α 4.1361.79 Pro15-β 1.83/1.16 28.34 Pro15-γ 1.58/1.48 24.83 Pro15-δ 3.79/3.5047.36 Pro15-C′ — 170.69  Phe16-NH 7.15 — Phe16-α 3.84 52.60 Phe16-β2.99/2.57 36.37 Phe16-γ — 139.47  Phe16-δ 7.25 129.33  Phe16-ε 7.24127.93  Phe16-ξ 7.13 125.65  Phe16-CH₂—OH 3.39/3.24 63.42 Phe16-CH₂—OH4.57 —

TABLE 3 NMR data (chemical shifts) for cephaibol A1 in DMSO at 300° K.Proton/carbon ¹H ¹³C Ac—Me 1.83 22.34 Ac—C′ — 170.18  Phe1-NH 8.22 —Phe1-α 4.39 54.88 Phe1-β 3.04/2.83 36.41 Phe1-γ — 137.53  Phe1-δ 7.29129.10  Phe1-ε 7.29 128.04  Phe1-ξ 7.21 126.32  Phe1-C′ — 172.09 Aib2-NH 8.50 — Aib2-α — 55.73 Aib2-β 1.31 24.39 Aib2-β′ 1.30 24.60Aib2-C′ — 174.55  Aib3-NH 7.73 — Aib3-α — 55.73 Aib3-β 1.32 25.41Aib3-β′ 1.29 23.12 Aib3-C′ — 175.38  Aib4-NH 7.80 — Aib4-α — 55.82Aib4-β 1.35 25.92 Aib4-β′ 1.34 23.12 Aib4-C′ — 175.60  Aib5-NH 7.66 —Aib5-α — 55.70 Aib5-β 1.40 26.24 Aib5-β′ 1.39 23.20 Aib5-C′ — 175.68 Ala6-NH 7.70 — Ala6-α 4.01 50.69 Ala6-β 1.38 16.47 Ala6-C′ — 174.55 Leu7-NH 7.67 — Leu7-α 3.96 53.49 Leu7-β 1.67/1.57 39.29 Leu7-γ 1.7224.18 Leu7-δ 0.90 22.58 Leu7-δ′ 0.84 21.29 Leu7-C′ — 171.81  Iva8-NH7.18 — Iva8-α — 59.29 Iva8-β 2.31/1.65 26.62 Iva8-γ 0.70  7.15 Iva8-αMe1.28 22.37 Iva8-C′ — 176.12  Aib9-NH 7.46 — Aib9-α — 56.16 Aib9-β 1.5023.12 Aib9-β′ 1.38 25.65 Aib9-C′ — 173.39  Hyp10-α 4.39 60.99 Hyp10-β2.17/1.78 36.95 Hyp10-γ 4.29 68.94 Hyp10-γOH 5.11 — Hyp10-δ 3.74/3.5556.27 Hyp10-C′ — 171.92  Gln11-NH 7.91 — Gln11-α 4.17 52.37 Gln11-β2.15/1.86 26.71 Gln11-γ 2.10 31.37 Gln11-δ — 173.08  Gln11-ε-NH₂7.19/6.70 — Gln11-C′ — 172.15  Iva12-NH 7.47 — Iva12-α — 58.47 Iva12-β2.15/1.78 28.04 Iva12-γ 0.74  6.94 Iva12-αMe 1.42 20.34 Iva12-C′ —172.82  Hyp13-α 4.54 60.54 Hyp13-β 2.17/1.78 37.33 Hyp13-γ 4.22 69.03Hyp13-γOH 5.08 — Hyp13-δ 3.68/3.38 56.45 Hyp13-C′ — 172.82  Aib14-NH7.95 — Aib14-α — 55.63 Aib14-β 1.39 23.50 Aib14-β′ 1.33 25.65 Aib14-C′ —171.71  Pro15-α 4.13 61.78 Pro15-β 1.85/1.17 28.33 Pro15-γ 1.59/1.4824.82 Pro15-δ 3.79/3.51 47.35 Pro15-C′ — 170.68  Phe16-NH 7.16 — Phe16-α3.84 52.59 Phe16-β 2.99/2.58 36.41 Phe16-γ — 139.46  Phe16-δ 7.25129.32  Phe16-ε 7.25 127.92  Phe16-ξ 7.14 125.64  Phe16-CH₂—OH 3.39/3.2563.41 Phe16-CH₂—OH 4.56 —

TABLE 4 NMR data (chemical shifts) for cephaibol B in DMSO at 300° K.Proton/carbon ¹H ¹³C Ac—Me 1.84 22.28 Ac—C′ — 170.37  Phe1-NH 8.28 —Phe1-α 4.33 55.14 Phe1-β 2.97/2.83 36.32 Phe1-γ — 137.36  Phe1-δ 7.29129.14  Phe1-ε 7.29 128.05  Phe1-ξ 7.22 126.37  Phe1-C′ — 172.40 Aib2-NH 8.58 — Aib2-α — 55.79 Aib2-β 1.28 23.69 Aib2-β′ 1.27 25.24Aib2-C′ — 174.81  Aib3-NH 7.62 — Aib3-α — 55.84 Aib3-β 1.32 24.10Aib3-β′ 1.29 24.60 Aib3-C′ — 175.10  Aib4-NH 7.73 — Aib4-α — 55.96Aib4-β 1.38 24.82 Aib4-β′ 1.38 24.82 Aib4-C′ — 175.45  Iva5-NH 7.49 —Iva5-α — 58.98 Iva5-β 1.99/1.70 ˜28.4(broad) Iva5-γ 0.80  7.49 Iva5-αMe1.35 21.23 Iva5-C′ — 175.56  Gly6-NH 7.95 — Gly6-α 3.77/3.64 43.34Gly6-C′ — 170.67  Leu7-NH 7.74 — Leu7-α 4.02 53.06 Leu7-β 1.69/1.5339.40 Leu7-γ 1.69 24.10 Leu7-δ 0.92 22.63 Leu7-δ′ 0.85 21.57 Leu7-C′ —171.71  Iva8-NH 7.41 — Iva8-α — 59.35 Iva8-β 2.17/1.67 27.89 Iva8-γ 0.74 7.32 Iva8-αMe 1.27 21.77 Iva8-C′ — 176.08  Aib9-NH 7.58 — Aib9-α —56.21 Aib9-β 1.48 23.15 Aib9-β′ 1.36 25.75 Aib9-C′ — 173.44  Hyp10-α4.39 60.99 Hyp10-β 2.16/1.78 36.83, 36.77^(a)) Hyp10-γ 4.28 68.94,68.84^(a)) Hyp10-γOH 5.11 — Hyp10-δ 3.73/3.50 56.09 Hyp10-C′ — 171.77 Gln11-NH 7.88 — Gln11-α 4.17 52.42 Gln11-β 2.15/1.87 26.66 Gln11-γ 2.1031.42 Gln11-δ — 173.00  Gln11-ε-NH₂ 7.18/6.70 — Gln11-C′ — 172.11 Iva12-NH 7.46 — Iva12-α — 58.47 Iva12-β 2.15/1.77 28.06 Iva12-γ 0.74 6.97 Iva12-αMe 1.42 20.35 Iva12-C′ — 172.81  Hyp13-α 4.53 60.55 Hyp13-β2.18/1.68 37.32, 37.26^(a)) Hyp13-γ 4.21 69.03, 68.94^(a)) Hyp13-γOH5.08 — Hyp13-δ 3.67/3.37 56.40 Hyp13-C′ — 172.81  Aib14-NH 7.94 —Aib14-α — 55.63 Aib14-β 1.40 23.49 Aib14-β′ 1.33 25.65 Aib14-C′ —171.71  Pro15-α 4.13 61.78 Pro15-β 1.83/1.16 28.33 Pro15-γ 1.58/1.4724.82 Pro15-δ 3.79/3.50 47.35 Pro15-C′ — 170.67  Phe16-NH 7.15 — Phe16-α3.83 52.59, 52.56^(a)) Phe16-β 2.99/2.57 36.37 Phe16-γ — 139.46  Phe16-δ7.25 129.32  Phe16-ε 7.25 127.91  Phe16-ξ 7.14 125.63  Phe16-CH₂—OH3.39/3.24 63.42, 63.30^(a)) Phe16-CH₂—OH 4.57 —

TABLE 5 NMR data (chemical shifts) for cephaibol C in DMSO at 300° K.Proton/carbon ¹H ¹³C Ac—Me 1.83 22.23 Ac—C′ — 170.27  Phe1-NH 8.27 —Phe1-α 4.34 54.99 Phe1-β 2.97/2.83 36.34 Phe1-γ — 137.35  Phe1-δ 7.29129.14  Phe1-ε 7.29 128.05  Phe1-ξ 7.22 126.38  Phe1-C′ — 172.39 Aib2-NH 8.58 — Aib2-α — 55.75 Aib2-β 1.28 23.51 Aib2-β′ 1.27 25.34Aib2-C′ — 174.81  Aib3-NH 7.63 — Aib3-α — 55.87 Aib3-β 1.32 23.97Aib3-β′ 1.29 24.55 Aib3-C′ — 175.08  Aib4-NH 7.69 — Aib4-α — 55.87Aib4-β 1.38 24.85 Aib4-β′ 1.38 24.55 Aib4-C′ — 175.62  Aib5-NH 7.56 —Aib5-α — 55.87 Aib5-β 1.41 24.85 Aib5-β′ 1.37 24.55 Aib5-C′ — 175.54 Gly6-NH 7.97 — Gly6-α 3.76/3.63 43.35 Gly6-C′ — 170.63  Leu7-NH 7.72 —Leu7-α 4.03 53.04 Leu7-β 1.69/1.51 39.41 Leu7-γ 1.69 24.11 Leu7-δ 0.9122.60 Leu7-δ′ 0.85 21.57 Leu7-C′ — 171.74  Iva8-NH 7.43 — Iva8-α — 59.34Iva8-β 2.17/1.69 27.87 Iva8-γ 0.74  7.31 Iva8-αMe 1.29 21.87 Iva8-C′ —176.04  Aib9-NH 7.54 — Aib9-α — 56.21 Aib9-β 1.48 23.23 Aib9-β′ 1.3625.49 Aib9-C′ — 173.59  Hyp10-α 4.39 61.12 Hyp10-β 2.16/1.77 36.75Hyp10-γ 4.20 68.93 Hyp10-γOH broad — Hyp10-δ 3.71/3.50 56.21 Hyp10-C′ —171.74  Gln11-NH 7.80 — Gln11-α 4.22 51.95 Gln11-β 2.18/1.83 26.17Gln11-γ 2.09 31.23 Gln11-δ — 173.10  Gln11-ε-NH₂ 7.18/6.69 — Gln11-C′ —172.02  Aib12-NH 7.75 — Aib12-α — 55.87 Aib12-β 1.50 22.80 Aib12-β′ 1.3825.73 Aib12-C′ — 172.26  Hyp13-α 4.51 60.54 Hyp13-β 2.16/1.69 37.22Hyp13-γ 4.21 68.99 Hyp13-γOH broad — Hyp13-δ 3.67/3.34 56.32 Hyp13-C′ —172.70  Aib14-NH 7.95 — Aib14-α — 55.65 Aib14-β 1.41 23.41 Aib14-β′ 1.3425.58 Aib14-C′ — 171.70  Pro15-α 4.13 61.77 Pro15-β 1.84/1.16 28.34Pro15-γ 1.58/1.49 24.55 Pro15-δ 3.80/3.51 47.36 Pro15-C′ — 170.63 Phe16-NH 7.14 — Phe16-α 3.83 52.46 Phe16-β 2.98/2.57 36.34 Phe16-γ —139.46  Phe16-δ 7.25 129.30  Phe16-ο 7.25 127.90  Phe16-ξ 7.13 125.63 Phe16-CH₂—OH 3.38/3.24 63.32 Phe16-CH₂—OH broad —

TABLE 6 NMR data (chemical shifts) for cephaibol D in DMSO at 300° K.Proton/carbon ¹H ¹³C Ac—Me 1.83 22.26 Ac—C′ — 170.27  Phe1-NH 8.28 —Phe1-α 4.34 55.05 Phe1-β 2.97/2.83 36.34 Phe1-γ — 137.35  Phe1-δ 7.29129.14  Phe1-ε 7.29 128.05  Phe1-ξ 7.22 126.38  Phe1-C′ — 172.40 Aib2-NH 8.59 — Aib2-α — 55.76 Aib2-β 1.28 23.47 Aib2-β′ 1.27 25.45Aib2-C′ — 174.89  Aib3-NH 7.62 — Aib3-α — 55.87 Aib3-β 1.32 23.98Aib3-β′ 1.29 24.84 Aib3-C′ — 175.04  Aib4-NH 7.69 — Aib4-α — 55.87Aib4-β 1.38 25.02 Aib4-β′ 1.37 24.48 Aib4-C′ — 175.59  Aib5-NH 7.57 —Aib5-α — 55.87 Aib5-β 1.40 24.84 Aib5-β′ 1.38 24.73 Aib5-C′ — 175.51 Gly6-NH 7.98 — Gly6-α 3.74/3.63 43.28 Gly6-C′ — 170.35  Leu7-NH 7.66 —Leu7-α 4.05 52.56 Leu7-β 1.66/1.52 39.25 Leu7-γ 1.66 24.07 Leu7-δ 0.9122.67 Leu7-δ′ 0.85 21.64 Leu7-C′ — 171.60  Aib8-NH 7.78 — Aib8-α — 56.15Aib8-β 1.45 25.45 Aib8-β′ 1.35 25.02 Aib8-C′ — 175.80  Aib9-NH 7.52 —Aib9-α — 56.22 Aib9-β 1.46 23.33 Aib9-β′ 1.34 25.45 Aib9-C′ — 173.64 Hyp10-α 4.38 61.09 Hyp10-β 2.15/1.78 36.58 Hyp10-γ 4.29 69.01 Hyp10-γOHbroad — Hyp10-δ 3.75/3.46 56.06 Hyp10-C′ — 171.67  Gln11-NH 7.77 —Gln11-α 4.22 51.96 Gln11-β 2.18/1.84 26.16 Gln11-γ 2.09 31.33 Gln11-δ —173.10  Gln11-ε-NH₂ 7.18/6.68 — Gln11-C′ — 172.03  Aib12-NH 7.76 —Aib12-α — 55.87 Aib12-β 1.50 23.81 Aib12-β′ 1.38 25.76 Aib12-C′ —172.27  Hyp13-α 4.51 60.54 Hyp13-β 2.16/1.70 37.25 Hyp13-γ 4.22 69.01Hyp13-γOH broad — Hyp13-δ 3.67/3.34 56.34 Hyp13-C′ — 172.75  Aib14-NH7.95 — Aib14-α — 55.66 Aib14-β 1.41 23.47 Aib14-β′ 1.34 25.60 Aib14-C′ —171.73  Pro15-α 4.13 61.77 Pro15-β  1.84/1.117 28.35 Pro15-γ 1.58/1.4924.84 Pro15-δ 3.80/3.52 47.36 Pro15-C′ — 170.69  Phe16-NH 7.14 — Phe16-α3.84 52.47 Phe16-β 2.98/2.58 36.34 Phe16-γ — 139.46  Phe16-δ 7.25129.31  Phe16-ε 7.25 127.90  Phe16-ξ 7.13 125.63  Phe16-CH₂—OH 3.39/3.2463.34 Phe16-CH₂—OH broad —

TABLE 7 NMR data (chemical shifts) for cephaibol E in DMSO at 300° K.Proton/carbon ¹H ¹³C Ac—Me 1.83 22.27 Ac—C′ — 170.36  Phe1-NH 8.28 —Phe1-α 4.34 55.10 Phe1-β 2.98/2.84 36.35 Phe1-γ — 137.36  Phe1-δ 7.29129.15  Phe1-ε 7.29 128.06  Phe1-ξ 7.23 126.39  Phe1-C′ — 172.42 Aib2-NH 8.58 — Aib2-α — 55.76 Aib2-β 1.28 25.39 Aib2-β′ 1.27 23.51Aib2-C′ — 174.90  Aib3-NH 7.62 — Aib3-α — 55.87 Aib3-β 1.32 24.08Aib3-β′ 1.29 24.63 Aib3-C′ — 175.06  Aib4-NH 7.69 — Aib4-α — 55.87Aib4-β 1.38 25.14 Aib4-β′ 1.38 24.94 Aib4-C′ — 175.60  Aib5-NH 7.58 —Aib5-α — 55.87 Aib5-β 1.40 24.94 Aib5-β′ 1.37 24.53 Aib5-C′ — 175.64 Gly6-NH 7.99 — Gly6-α 3.74/3.64 43.35 Gly6-C′ — 170.43  Leu7-NH 7.68 —Leu7-α 4.04 52.73 Leu7-β 1.67/1.54 39.24 Leu7-γ 1.67 24.08 Leu7-δ 0.9122.67 Leu7-δ′ 0.86 21.64 Leu7-C′ — 171.71  Aib8-NH 7.72 — Aib8-α — 56.18Aib8-β 1.45 25.26 Aib8-β′ 1.34 25.66 Aib8-C′ — 175.84  Aib9-NH 7.54 —Aib9-α — 56.22 Aib9-β 1.46 23.26 Aib9-β′ 1.34 25.66 Aib9-C′ — 173.48 Hyp10-α 4.38 60.95 Hyp10-β 2.16/1.79 36.71 Hyp10-γ 4.29 68.95 Hyp10-γOHbroad — Hyp10-δ 3.77/3.48 56.00 Hyp10-C′ — 171.75  Gln11-NH 7.85 —Gln11-α 4.17 52.48 Gln11-β 2.16/1.89 26.63 Gln11-γ 2.11 31.49 Gln11-δ —173.08  Gln11-ε-NH₂ 7.19/6.69 — Gln11-C′ — 172.13  Iva12-NH 7.47 —Iva12-α — 58.47 Iva12-β 2.15/1.78 28.09 Iva12-γ 0.75  7.01 Iva12-αMe1.41 20.37 Iva12-C′ — 172.83  Hyp13-α 4.53 60.54 Hyp13-β 2.17/1.68 37.32Hyp13-γ 4.21 69.00 Hyp13-γOH broad — Hyp13-δ 3.69/3.38 56.45 Hyp13-C′ —172.83  Aib14-NH 7.94 — Aib14-α — 55.64 Aib14-β 1.39 23.51 Aib14-β′ 1.3325.66 Aib14-C′ — 171.71  Pro15-α 4.13 61.78 Pro15-β 1.85/1.17 28.33Pro15-γ 1.59/1.48 24.82 Pro15-δ 3.80/3.51 47.36 Pro15-C′ — 170.68 Phe16-NH 7.15 — Phe16-α 3.84 52.58 Phe16-β 2.98/2.58 36.35 Phe16-γ —139.47  Phe16-δ 7.26 129.33  Phe16-ε 7.26 127.92  Phe16-ξ 7.15 125.64 Phe16-CH₂—OH 3.39/3.25 63.38 Phe16-CH₂—OH broad —

TABLE 8 NMR data (chemical shifts) for cephaibol P in DMSO at 300° K.Proton ¹H Ac—Me 1.83 Ac—C′ — Phe1-NH 8.23 Phe1-α 4.48 Phe1-β 3.05/2.84Phe1-δ 7.27 Phe1-ε 7.27 Phe1-ξ 7.19 Iva2-NH 8.28 Iva2-β 1.86/1.66 Iva2-γ0.74 Iva2-αMe 1.27 Gln3-NH 8.14 Gln3-α 3.97 Gln3-β 1.88 Gln3-γ 2.18Gln3-ε-NH₂ 7.34/6.88 Aib4-NH 7.99 Aib4-β 1.39 Aib4-β′ 1.35 Ile5-NH 7.53Ile5-α 3.88 Ile5-β 1.88 Ile5-γMe 0.84 Ile5-γ 1.45/1.18 Ile5-δ 0.78Thr6-NH 7.60 Thr6-α 3.82 Thr6-β 4.09 Thr6-γ 1.12 Thr6-OH 4.96 Aib7-NH7.77 Aib7-β 1.36 Aib7-β′ 1.36 Leu8-NH 7.26 Leu8-α 4.21 Leu8-β 1.65/1.56Leu8-γ 1.68 Leu8-δ 0.85 Leu8-δ′ 0.78 Aib9-NH 7.94 Aib9-β 1.52 Aib9-β′1.42 Hyp10-α 4.39 Hyp10-β 2.16/1.77 Hyp10-γ 4.26 Hyp10-γOH 5.12 Hyp10-δ3.76/3.44 Gln11-NH 7.85 Gln11-α 4.22 Gln11-β 1.89 Gln11-γ 2.14/2.12Gln11-ε-NH₂ 7.20/6.70 Aib12-NH 7.79 Aib12-β 1.51 Aib12-β′ 1.39 Hyp13-α4.53 Hyp13-β 2.18/1.71 Hyp13-γ 4.22 Hyp13-γOH 5.09 Hyp13-δ 3.67/3.38Aib14-NH 8.07 Aib14-β 1.47 Aib14-β′ 1.38 Pro15-α 4.17 Pro15-β 1.90/1.04Pro15-γ 1.63/1.51 Pro15-δ 3.87/3.58 Pro15-C′ — Phe16-NH 7.67 Phe16-α4.35 Phe16-β 3.26/2.80 Phe16-δ 7.32 Phe16-ε 7.26 Phe16-ξ 7.19 Ser17-NH7.39 Ser17-α 4.24 Ser17-β 3.71 Ser17-βOH 4.77(broad)

TABLE 9 NMR data (chemical shifts) for cephaibol Q in DMSO at 300° K.Proton ¹H Ac—Me 1.83 Ac—C′ — Phe1-NH 8.23 Phe1-α 4.48 Phe1-β 3.05/2.84Phe1-δ 7.27 Phe1-ε 7.27 Phe1-ξ 7.19 Iva2-NH 8.28 Iva2-β 1.86/1.66 Iva2-γ0.74 Iva2-αMe 1.27 Gln3-NH 8.14 Gln3-α 3.97 Gln3-β 1.88 Gln3-γ 2.18Gln3-ε-NH₂ 7.34/6.88 Aib4-NH 7.99 Aib4-β 1.39 Aib4-β′ 1.35 Ile5-NH 7.53Ile5-α 3.88 Ile5-β 1.88 Ile5-γMe 0.84 Ile5-γ 1.45/1.18 Ile5-δ 0.78Thr6-NH 7.60 Thr6-α 3.83 Thr6-β 4.09 Thr6-γ 1.11 Thr6-OH 4.96 Aib7-NH7.74 Aib7-β 1.36 Aib7-β′ 1.36 Leu8-NH 7.28 Leu8-α 4.16 Leu8-β 1.65/1.56Leu8-γ 1.69 Leu8-δ 0.85 Leu8-δ′ 0.78 Aib9-NH 7.97 Aib9-β 1.50 Aib9-β′1.42 Pro10-α 4.30 Pro10-β 2.22/1.69 Pro10-γ 1.84 Pro10-δ 3.75/3.53Gln11-NH 7.79 Gln11-α 4.24 Gln11-β 1.88 Gln11-γ 2.14/2.12 Gln11-ε-NH₂7.19/6.71 Aib12-NH 7.78 Aib12-β 1.50 Aib12-β′ 1.39 Hyp13-α 4.53 Hyp13-β2.18/1.71 Hyp13-γ 4.22 Hyp13-γOH 5.09 Hyp13-δ 3.67/3.38 Aib14-NH 8.07Aib14-β 1.47 Aib14-β′ 1.38 Pro15-α 4.17 Pro15-β 1.90/1.04 Pro15-γ1.63/1.51 Pro15-δ 3.87/3.58 Pro15-C′ — Phe16-NH 7.67 Phe16-α 4.35Phe16-β 3.26/2.80 Phe16-δ 7.32 Phe16-ε 7.26 Phe16-ξ 7.19 Ser17-NH 7.39Ser17-α 4.24 Ser17-β 3.71 Ser17-βOH 4.77(broad)

Example 10 Determination of Anthelmintic Action

For the testing of the action of the cephaibols on helminths, an invitro test (larval development test) was carried out with larvae of thechicken roundworm Ascaridia galli. Embryonate eggs of A. galli weresurface-sterilized by treatment with 5% strength sodium hypochloritesolution and, after removal of this solution, mechanically opened bymeans of rotating glass beads (5 mm). The hatched L2 larvae wereenriched via larval enrichment processes, taken up in nutrient mediumand incubated at 41° C. and 10% CO₂.

48 hours after hatching, the larvae were incubated (41° C., 10% CO₂) inmicrotiter plates (96 well) for 5 days with medicated medium (200 μl) inthe concentrations 200, 100, 50 . . . 0.1 μg/ml. During the 5-dayincubation period, motility, morphology, and vitality were checkedmicroscopically and documented on a daily basis. After medication, thelarval culture was incubated for 48 hours with Neutral Red in aconcentration of 0.16%; after removal of the Neutral Red by change ofmedium, the enrichment of the vital dye in the intestine of the larvaewas assessed as an index for active food intake and thus as proof of thevitality. The medication of the larvae with cephaibol A in theconcentrations 200, 100, 50, and 25 μg/ml led to 100% mortality; at aconcentration of 12.5 μg/ml and 6.25 μg/ml, the mortality was 92% and20% respectively.

Example 11 Action on Ectoparasites

The action against ectoparasites was checked in the flea larval test(cat flea=Ctenocephalides felis). 5 mg of cephaibol A were dissolved in0.5 ml of acetone and mixed into 500 mg of blood meal (defibrinatedsheep blood). After the solvent had evaporated, 2 g of quartz sand weremixed with medicated blood meal such that preparation concentrations of2000, 1000, 500, 250 ppm etc. resulted. 15 flea eggs per medicatedsample or solvent control were added to the mixture of medicated bloodmeal and quartz sand, which was then incubated at 37° C. and highatmospheric humidity. The larval development, pupation and developmentto the adult stage were checked at an interval of 3-5 days and themortality rates were documented.

Depending on the dose, a larvicidal action of >90% in comparison withthe solvent control was observed.

Example 12 Determination of the Antimicrobial Action

Table 10 shows some minimal inhibitory concentrations (MIC) of theantimicrobial spectrum.

TABLE 10 The minimal inhibitory concentrations (MIC) of cephaibol Aagainst some selected microorganisms. MIC Strain [μg/ml] Enterococcushirae ATCC 10541 64 Enterococcus faecalis ATCC 29212 64 Enterobactercloacae P 99 >128 Enterobacter cloacae 1321 E >128 Escherichia coliTEM >128 Escherichia coli 1507 E >128 Escherichia coli DC 0 >128Escherichia coli DC 2 >128 Escherichia coli ATCC 25922 >128 Klebsiellaaerogenes 1082 E >128 Klebsiella oxytoca 1522 E >128 Pseudomonasaeruginosa 77/2 >128 Pseudomonas aeruginosa 1771 >128 Pseudomonasaeruginosa 1771 M >128 Pseudomonas aeruginosa ATCC 9027 >128 Pseudomonasaeruginosa ATCC 27853 >128 Staphylococcus aureus 285 8 Staphylococcusaureus ATCC 29213 16 Streptococcus pyogenes 308 A 32 Streptococcuspyogenes 77 A >128 Bordetella bronchiseptica ATCC 4617 >128 Pasteurellamultocida 3; tox. pos. >128 Pasteurella haemolytica 7 >128 Bordetellabronchiseptica 9 >128 Bordetella bronchiseptica 10 >128 Pasteurellamultocida P 48 >128 Pasteurella haemolytica P 53 >128 Mycoplasmagallisepticum 64 Mycoplasma mycoides LC 128 Mycoplasma mycoides LC 64

10 1 16 PRT Artificial Sequence Description of Artificial SequenceSynthetic peptide 1 Xaa Xaa Xaa Xaa Xaa Gly Leu Xaa Xaa Xaa Gln Xaa XaaXaa Pro Xaa 1 5 10 15 2 16 PRT Artificial Sequence Description ofArtificial Sequence Synthetic peptide 2 Xaa Xaa Xaa Xaa Xaa Gly Leu XaaXaa Xaa Gln Xaa Xaa Xaa Pro Xaa 1 5 10 15 3 16 PRT Artificial SequenceDescription of Artificial Sequence Synthetic peptide 3 Xaa Xaa Xaa XaaXaa Gly Leu Xaa Xaa Xaa Gln Xaa Xaa Xaa Pro Xaa 1 5 10 15 4 16 PRTArtificial Sequence Description of Artificial Sequence Synthetic peptide4 Xaa Xaa Xaa Xaa Xaa Gly Leu Xaa Xaa Xaa Gln Xaa Xaa Xaa Pro Xaa 1 5 1015 5 16 PRT Artificial Sequence Description of Artificial SequenceSynthetic peptide 5 Xaa Xaa Xaa Xaa Xaa Gly Leu Xaa Xaa Xaa Gln Xaa XaaXaa Pro Xaa 1 5 10 15 6 17 PRT Artificial Sequence Description ofArtificial Sequence Synthetic peptide 6 Xaa Xaa Gln Xaa Ile Thr Xaa LeuXaa Xaa Gln Xaa Xaa Xaa Pro Phe 1 5 10 15 Ser 7 17 PRT ArtificialSequence Description of Artificial Sequence Synthetic peptide 7 Xaa XaaGln Xaa Ile Thr Xaa Leu Xaa Pro Gln Xaa Xaa Xaa Pro Phe 1 5 10 15 Ser 816 PRT Artificial Sequence Description of Artificial Sequence Syntheticpeptide 8 Xaa Xaa Xaa Xaa Xaa Ala Leu Xaa Xaa Xaa Gln Xaa Xaa Xaa ProXaa 1 5 10 15 9 16 PRT Artificial Sequence Description of ArtificialSequence Synthetic peptide 9 Xaa Xaa Xaa Xaa Xaa Xaa Leu Xaa Xaa Xaa GlnXaa Xaa Xaa Pro Xaa 1 5 10 15 10 17 PRT Artificial Sequence Descriptionof Artificial Sequence Synthetic peptide 10 Xaa Xaa Gln Xaa Ile Thr XaaLeu Xaa Xaa Gln Xaa Xaa Xaa Pro Phe 1 5 10 15 Ser

We claim:
 1. An isolated compound of the formula I (SEQ ID NO:9)AcPhe-Aib-Aib-Aib-x-w-Leu-y-Aib-Hyp-Gln-z-Hyp-Aib-Pro-R  (I) Wherein Ris Phe-ol or Phe-al and w, x, y, and z have the following meanings: a) wis Gly or Ala; x is Aib and y and z are Iva; b) w is Gly; x, y and z areIva; c) w is Gly; x and z are Aib and y is Iva; d) w is Gly; x, y and zare Aib; or e) w is Gly; x and y are Aib, and z is Iva; or aphysiologically tolerable salt thereof.
 2. The compound of the formula Ias claimed in claim 1, wherein R is Phe-ol, or a physiologicallytolerable salt thereof.
 3. An isolated compound of the formula II (SEQID NO:10)AcPhe-Iva-Gln-Aib-Ile-Thr-Aib-Leu-Aib-x-Gln-Aib-Hyp-Aib-Pro-Phe-Ser  (I)Wherein x is Hyp or Pro, or a physioloically tolerable salt thereof. 4.An isolated compound selected fromAcPhe-Aib-Aib-Aib-Aib-Gly-Leu-Iva-Aib-Hyp-Gln-Iva-Hyp-Aib-Pro-Phe-ol(SEQ ID NO:1);AcPhe-Aib-Aib-Aib-Aib-Ala-Leu-Iva-Aib-Hyp-Gln-Iva-Hyp-Aib-Pro-Phe-ol(SEQ ID NO:8);AcPhe-Aib-Aib-Aib-Iva-Gly-Leu-Iva-Aib-Hyp-Gln-Iva-Hyp-Aib-Pro-Phe-ol(SEQ ID NO:2);AcPhe-Aib-Aib-Aib-Aib-Gly-Leu-Iva-Aib-Hyp-Gln-Aib-Hyp-Aib-Pro-Phe-ol(SEQ ID NO:3);AcPhe-Aib-Aib-Aib-Aib-Gly-Leu-Aib-Aib-Hyp-Gln-Aib-Hyp-Aib-Pro-Phe-ol(SEQ ID NO:4);AcPhe-Aib-Aib-Aib-Aib-Gly-Leu-Aib-Aib-Hyp-Gln-Iva-Hyp-Aib-Pro-Phe-ol(SEQ ID NO:5);AcPhe-Iva-Gln-Aib-Ile-Thr-Aib-Leu-Aib-Hyp-Gln-Aib-Hyp-Aib-Pro-Phe-Ser(SEQ ID NO:6); orAcPhe-Iva-Gln-Aib-Ile-Thr-Aib-Leu-Aib-Pro-Gln-Aib-Hyp-Aib-Pro-Phe-Ser(SEQ ID NO:7); or a physiologically tolerable salt thereof.
 5. A processfor the preparation of a compound of the formula I (SEQ ID NO:9) or II(SEQ ID NO:10), or a physiologically tolerable salt thereof, as claimedin claim 1 or claim 3, which comprises fermenting the microorganismAcremonium tubakii FH 1685 DSM 12774 under suitable conditions in aculture medium until one or more compounds of the formula I or IIaccumulate in the culture medium and then isolating them from theculture medium and, optionally converting them into physiologicallytolerable salts.
 6. The process as claimed in claim 5, wherein thefermentation is carried out under aerobic conditions at a temperaturefrom about 18° C. to about 35° C. and at a pH from about 6 to about 8.7. A pharmaceutical composition comprising at least one compound of theformula I (SEQ ID NO:9) or II (SEQ ID NO:10), or a physiologicallytolerable salt thereof, as claimed in claim 1 or claim 3 and at leastone of a suitable carrier, an excipient, and a vehicle foradministration.
 8. A process for the preparation of a pharmaceuticalcomposition as claimed in claim 7, which comprises bringing at least onecompound of the formula I (SEQ ID NO:9) or II (SEQ ID NO:10), or aphysiologically tolerable salt thereof, into a suitable administrationform.